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A wheat gluten fermentation process with the inoculation of different microorganisms under salt-free conditions has the potential to produce varying flavour profiles. As research on the co-fermentation of yeasts and lactic acid bacteria (LAB) in salt-free wheat gluten fermentation is scarce, the current work studied the flavour impact on fermented wheat gluten by the co-inoculation of Latilactobacillus sakei with one yeast (Saccharomyces boulardii or Pichia kluyveri). The results showed that similar glucose and organic acid levels were detected, but early death of yeasts was observed during liquid-state fermentation (LSF) in co-fermentations. The concentrations of most free amino acids were comparable. Volatile compound analysis showed synergistic effects in co-cultured fermentations on the production of certain compounds such as isoamyl acetate. Principal component analysis revealed clear differences in volatile profiles between co-fermentation and single-strain fermentation. Therefore, a fermented sauce produced by co-inoculating LAB and yeast with a new and fruitier flavour was developed.

Liquid-state fermentation (LSF) and solid-state fermentation (SSF) are two forms of industrial production of lactic acid bacteria (LAB). The choice of two fermentations for LAB production has drawn wide concern. In this study, the tolerance of bacteria produced by the two fermentation methods to acid stress was compared, and the reasons for the tolerance differences were analyzed at the physiological and transcriptional levels. The survival rate of the bacterial agent obtained from solid-state fermentation was significantly higher than that of bacteria obtained from liquid-state fermentation after spray drying and cold air drying. However, the tolerance of bacterial cells obtained from liquid-state fermentation to acid stress was significantly higher than that from solid-state fermentation. The analysis at physiological level indicated that under acid stress, cells from liquid-state fermentation displayed a more solid and complete membrane structure, higher cell membrane saturated fatty acid, more stable intracellular pH, and more stable activity of ATPase and glutathione reductase, compared with cells from solid-state fermentation, and these physiological differences led to better tolerance to acid stress. In addition, transcriptomic analysis showed that in the cells cultured from liquid-state fermentation, the genes related to glycolysis, inositol phosphate metabolism, and carbohydrate transport were down-regulated, whereas the genes related to fatty acid synthesis and glutamate metabolism were upregulated, compared with those in cells from solid-state fermentation. In addition, some genes related to acid stress response such as cspA, rimP, rbfA, mazF, and nagB were up-regulated. These findings provide a new perspective for the study of acid stress tolerance of L. paracasei Zhang and offer a reference for the selection of fermentation methods of LAB production.

Liquid-state fermentation has been increasingly applied in the industrial glutinous rice wine (GRW) production. However, products brewed by this emerging technique possess some deficiencies in flavor quality. Therefore, this study firstly developed and optimized an innovative pulping technique by the synchronously pulping and gelatinizing treatment (Process I) to improve GRW flavor quality, and then revealed the influences of Process I on raw materials properties, oenological parameters, fermentation process, and flavor characteristics of GRW. Results show that Process I significantly (p < 0.05) enriched the soluble solid and crude protein content of glutinous rice milk by improving gelatinization degree and pulping efficiency, which consequently enhanced the microbial growth, glycolysis, and protein decomposition during the GRW fermentation process. GC–MS analysis shows that Process I sequentially significantly (p < 0.05) enhanced the esterification and Ehrlich or Harrison pathway during the fermentation process. This contributed to a higher content of key ester and alcohol compounds.

The property differences between bacteria produced from solid-state and liquid-state fermentations have always been the focus of attention. This study analyzed the stress tolerance and transcriptomic differences of the probiotic Lacticaseibacillus casei Zhang produced from solid-state and liquid-state fermentations under no direct stress. The total biomass of L. casei Zhang generated from liquid-state fermentation with MRS medium (LSF-MRS) was 2.24 times as much as that from solid-state fermentation with soybean meal-wheat bran (SSF-SW) medium. Interestingly, NaCl, H2O2, and ethanol stress tolerances and the survival rate after L. casei Zhang agent preparation from SSF-SW fermentation were significantly higher than those from LSF-MRS fermentation. The global transcriptomic analysis revealed that in L. casei Zhang produced from SSF-SW fermentation, carbohydrate transport, gluconeogenesis, inositol phosphate metabolism were promoted, that pentose phosphate pathway was up-regulated to produce more NADPH, that citrate transport and fermentation was extremely significantly promoted to produce pyruvate and ATP, and that pyruvate metabolism was widely up-regulated to form lactate, acetate, ethanol, and succinate from pyruvate and acetyl-CoA, whereas glycolysis was suppressed, and fatty acid biosynthesis was suppressed. Moreover, in response to adverse stresses, some genes encoding aquaporins (GlpF), superoxide dismutase (SOD), nitroreductase, iron homeostasis-related proteins, trehalose operon repressor TreR, alcohol dehydrogenase (ADH), and TetR/AcrR family transcriptional regulators were up-regulated in L. casei Zhang produced from SSF-SW fermentation. Our findings provide novel insight into the differences in growth performance, carbon and lipid metabolisms, and stress tolerance between L. casei Zhang from solid-state and liquid-state fermentations.

To expand the range of soluble carbon sources for enzyme production by the microbial system, we investigated the ability of different sugars in cellulase production. Carbon sources play a vital role in cell metabolism and the synthesis of cellulase. Although the insoluble cellulosic materials are considered the most effective natural inducers for cellulase production by microorganisms in terms of both enzyme yield and productivity, their insolubility causes many problems and presents a major drawback that is partly responsible for the high cost of cellulase production. Mostly, the insolubility of sugar polymer leads to difficult and complex fermentation operations, including sterilization, cell biomass measurement, mixing and aeration of the fermentation broth, continuous feeding/sampling, and subsequent enzyme purification. Second, cellulase gets absorbed into the solid cellulose surface, leading to enzyme loss. The present paper gives a comparative view on the utility of pure sugars (lactose, CMC, sucrose, maltose, cellobiose, xylose, trehalose, arabinose) and sugar alcohols (sorbitol, mannitol) on cellulase production by various fungal strains. It also describes the blending effect of sugars as well as sugar alcohols on cellulase production by fungal strains.

Red pigment is one of natural coloring agents produced from the secondary metabolites of Monascus purpureus . Measures are taken to increase the pigment production. Effect of the extremely low-frequency magnetic field on the red and yellow pigment production of M. purpureus in liquid-state fermentation was estimated by exposing fermentation broth, medium and sterile distilled water used to prepare medium. The maximum yield of red and yellow pigment production had about 20 and 36 % increase for 8 days at 0.4 mT when treating fermentation broth compared to the control value. For sterile distilled water, a significant increase occurred at 0.4 mT compared to 0.1 mT for 5 h. However, the red and yellow pigment production decreased significantly at magnetic fields of 0.9 and 1.2 mT when treating fermentation broth and sterile distilled water. In contrast, the yield of both red and yellow pigment production decreased significantly compared with the control exposure for 3 and 5 h at a magnetic field of 0.4 mT. Different solution (cellular suspension, medium and sterile distilled water) exposure to magnetic fields has different bioeffect on M. purpureus SKY219. The appropriate magnetic field treatment could increase the efficiency of red and yellow pigment production.

Entomopathogenic fungi (EPF) can be defined as beneficial multifunctional eukaryotic microorganisms that display pivotal ecological services in pest management, with some species possessing the special ability to establish mutualistic relationships with plants. Mass production of these fungi is critical to support affordable widespread commercialization and worldwide field application. Among the mass production methods explored mainly by industry, submerged liquid fermentation is a robust and versatile technology that allows the formation of different types of propagules designated for various applications in pest control. Many hypocrealean EPF are easily culturable on artificial substrates by producing single-celled structures (hyphal bodies, blastospores, and submerged conidia) or multicellular structures (mycelium and microsclerotia). Less frequently, some EPF may form environmentally resistant chlamydospores, but these structures have almost always been overlooked. A continued research pipeline encompassing screening fungal strains, media optimization, and proper formulation techniques aligned with the understanding of molecular cues involved in the formation and storage stability of these propagules is imperative to unlock the full potential and to fine-tune the development of robust and effective biocontrol agents against arthropod pests and vectors of diseases. Finally, we envision a bright future for the submerged liquid fermentation technology to supplement or replace the traditional solid substrate fermentation method for the mass production of many important EPF. 653 __ |a Biological control; Mass production; Blastospores; Microsclerotia; Submerged conidia; Bioreactor.

Baijiu is the national liquor of China and the world’s most consumed spirit, which is produced using a unique and traditional solid-state fermentation (SSF) process. The development of an automatic and intelligent technology for SSF is more difficult than that for liquid-state fermentation. However, the technological upgrading of the SSF process is crucial for reducing the labor intensity, saving manpower, avoiding the waste of materials and energy, and providing a favorable operation environment for workers; moreover, it provides a great reference value to similar industries. This article reviews the latest application progresses in automatic and intelligent technologies for Baijiu production. The important technical processes are introduced successively, including the production of Jiuqu, SSF, solid-state distillation, storage, and blending. The bottlenecks and challenges are pointed out for automatic and intelligent upgrading of these technical processes. Furthermore, the typical technology application cases in an integrated automatic production line of Baijiu are also summarized. Next, the industrial development status of Baijiu production is compared with those of other liquors in the world. Finally, future development directions are proposed. This review will provide an overall introduction and objective analysis of recent developments and current challenges in Baijiu manufacturing so as to promote the intelligent brewing of Baijiu.

This study was to establish an efficient strategy based on inoculum-morphology control for the submerged mycelial fermentation of an edible and medicinal fungus, Inonotus hispidus . Two major morphological forms of the mycelial inoculum were compared, dispersed mycelial fragments versus aggregated mycelial clumps. The dispersed one was more favorable for the fermentation, starting with a shorter lag period and attaining a higher biomass yield and more uniform mycelium pellets in shake flasks. The mycelial pellets taken from the shake flask culture on day 6 were fragmented at 26,000 rpm in a homogenizer, and a shear time of 3 min provided the optimal inoculum. The inoculum and culture conditions were further verified in 5-L stirred tank fermenters and then the fermentation was scaled-up in a 100-L stirred tank. With the optimized inoculum and process conditions plus a fed-batch operation, much higher productivities, including 22.23 g/L biomass, 3.31 g/L EPS, and 5.21 g/L IPS, were achieved in the 100-L fermenter than in the flask culture. A composition analysis showed that the I. hispidus mycelium produced by the fermentation was rich in protein, dietary fiber, and polysaccharides which may be beneficial to health. Overall, the results have shown that the inoculum characteristics including age, morphology, and state of aggregation have significant impact on the productivity of mycelial biomass and polysaccharides in a submerged mycelial fermentation of the I. hispidus fungus. Graphical Abstract (created with BioRender.com)

Oxygen is crucial for converting glucose to gluconic acid catalyzed by glucose oxidase (Gox). However, industrial gluconic acid production faces oxygen supply limitations. To enhance Gox efficiency, Vitreoscilla hemoglobin (VHb) has been considered as an efficient oxygen transfer carrier. This study identified GoxA, a specific isoform of Gox in the industrial gluconic acid-producing strain of Aspergillus niger . Various forms of VHb expression in A. niger were tested to improve GoxA’s catalytic efficiency. Surprisingly, the expression of free VHb, both intracellularly and extracellularly, did not promote gluconic acid production during shake flask fermentation. Then, five fusion proteins were constructed by linking Gox and VHb using various methods. Among these, VHb-GS1-GoxA, where VHb’s C-terminus connected to GoxA’s N-terminus via the flexible linker GS1, demonstrated a significantly higher Kcat/Km value (96% higher) than GoxA. Unfortunately, the expression of VHb-GS1-GoxA in A. niger was limited, resulting in a low gluconic acid production of 3.0 g/L. To overcome the low expression problem, single- and dual-strain systems were designed with tools of SpyCatcher/SpyTag and SnoopCatcher/SnoopTag. In these systems, Gox and VHb were separately expressed and then self-assembled into complex proteins. Impressively, the single-strain system outperformed the GoxA overexpression strain S1971, resulting in 23% and 9% higher gluconic acid production under 0.6 vvm and 1.2 vvm aeration conditions in the bioreactor fermentation, respectively. The successful construction of Gox and VHb fusion or complex proteins, as proposed in this study, presents promising approaches to enhance Gox catalytic efficiency and lower aerodynamic costs in gluconic acid production. Key points • Overexpressing free VHb in A. niger did not improve the catalytic efficiency of Gox • The VHb-GS1-GoxA showed an increased Kcat/Km value by 96% than GoxA • The single-strain system worked better in the gluconic acid bioreactor fermentation